anti cyr61  (Proteintech)

Journal: Journal of Dental Sciences

Article Title: Laser-modified titanium surfaces induce sex-dimorphic secretion of angiogenic factors by gingiva-derived mesenchymal stromal cells

doi: 10.1016/j.jds.2025.09.013

Figure Lengend Snippet: Laser-modified titanium upregulates angiogenesis-related genes in human gingiva-derived mesenchymal stromal cells (GMSCs). Assay for transposase-accessible chromatin sequencing (ATAC-seq) and RNA-sequencing (RNA-seq) were performed on GMSCs cultured on machined, lasered, and SLA (sand-blasted, large-grit, acid-etched) titanium discs for 72 h (n = 2 per group; age-matched 50-year-old female and male). (A) Venn diagrams show differentially upregulated (UP) and downregulated (DOWN) genes in the lasered group and SLA group compared to the machined group, respectively. (B) Volcano plot highlights differentially expressed angiogenesis-related genes. (C) Gene Ontology (GO) enrichment analysis of genes upregulated in the lasered group versus the machined group. (D) Thirty-three genes were identified at the intersection of GO:0016477 (cell migration) and genes upregulated in both female and male GMSCs on laser-modified versus machined surfaces. The heatmap shows shared migration-related genes upregulated in both sexes. (E) Chromatin accessibility and RNA expression levels of CCN1 and EDIL3. F, female. M, male. CCN1, cellular communication network factor 1. EDIL3, EGF like repeats and discoidin domains 3.

Article Snippet: Subsequently, the medium was replaced with fresh exosome-free α-MEM and cells were cultured for an additional 48 h. Conditioned medium was collected for the tube formation assay, and GMSCs were harvested for RNA extraction. mRNA expression was validated using TaqMan Fast Advanced Master Mix (4444556, Invitrogen) with TaqMan Gene Expression Assays (Invitrogen) for CYR61 (Hs00155479_m1), EDIL3 (Hs00964112_m1), and GAPDH (Hs02786624_g1), according to the manufacturer's protocol.

Techniques: Modification, Derivative Assay, Sequencing, RNA Sequencing, Cell Culture, Migration, RNA Expression

Journal: Journal of Dental Sciences

Article Title: Laser-modified titanium surfaces induce sex-dimorphic secretion of angiogenic factors by gingiva-derived mesenchymal stromal cells

doi: 10.1016/j.jds.2025.09.013

Figure Lengend Snippet: Laser-modified titanium surfaces upregulate CCN1 expression in human gingiva-derived mesenchymal stromal cells (GMSCs). Relative gene expression levels of ADM2, CCN1, EDIL3, and β-actin in GMSCs cultured on three different titanium disc surfaces: machined, lasered, and SLA (sand-blasted, large-grit, acid-etched). (A) CCN1 mRNA levels were significantly upregulated in the laser-treated and SLA groups compared to the machined group. (B) and (C) No significant differences in ADM2 and EDIL3 expression were observed among the three groups. (D) β-actin expression was significantly increased in the laser-treated and SLA groups compared to the machined group. Data are means ± standard deviation (n = 8 per group, 4 females and 4 males). Paired t-test. ∗ P < 0.05, ∗∗∗∗ P < 0.0001. mRNA, messenger ribonucleic acid. CCN1, cellular communication network factor 1. ADM2, adrenomedullin 2. EDIL3, EGF like repeats and discoidin domains 3. RPS18, ribosomal protein S18.

Article Snippet: Subsequently, the medium was replaced with fresh exosome-free α-MEM and cells were cultured for an additional 48 h. Conditioned medium was collected for the tube formation assay, and GMSCs were harvested for RNA extraction. mRNA expression was validated using TaqMan Fast Advanced Master Mix (4444556, Invitrogen) with TaqMan Gene Expression Assays (Invitrogen) for CYR61 (Hs00155479_m1), EDIL3 (Hs00964112_m1), and GAPDH (Hs02786624_g1), according to the manufacturer's protocol.

Techniques: Modification, Expressing, Derivative Assay, Gene Expression, Cell Culture, Standard Deviation

Journal: Journal of Dental Sciences

Article Title: Laser-modified titanium surfaces induce sex-dimorphic secretion of angiogenic factors by gingiva-derived mesenchymal stromal cells

doi: 10.1016/j.jds.2025.09.013

Figure Lengend Snippet: Sex-dimorphic secretion of EDIL3 and CCN1 by human gingiva-derived mesenchymal stromal cells (GMSCs). ELISA assays were performed to quantify CCN1 and EDIL3 levels in conditioned medium collected from GMSCs cultured on machined, lasered, and SLA (sand-blasted, large-grit, acid-etched) titanium discs for 72 h. (A) ELISA results from conditioned medium without Triton X treatment. Data are mean ± standard deviation (n = 8; four females and four males). Dotted line: paired t-test. Solid line: unpaired t-test. ∗ P < 0.05. (B) ELISA results from conditioned medium treated with Triton X. Data are mean ± standard deviation (n = 8; four females and four males). Dotted line: paired t-test. Solid line: unpaired t-test. ∗ P < 0.05, ∗∗ P < 0.01. F, female. M, male. CCN1, cellular communication network factor 1. EDIL3, EGF like repeats and discoidin domains 3.

Article Snippet: Subsequently, the medium was replaced with fresh exosome-free α-MEM and cells were cultured for an additional 48 h. Conditioned medium was collected for the tube formation assay, and GMSCs were harvested for RNA extraction. mRNA expression was validated using TaqMan Fast Advanced Master Mix (4444556, Invitrogen) with TaqMan Gene Expression Assays (Invitrogen) for CYR61 (Hs00155479_m1), EDIL3 (Hs00964112_m1), and GAPDH (Hs02786624_g1), according to the manufacturer's protocol.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Standard Deviation

Journal: Journal of Dental Sciences

Article Title: Laser-modified titanium surfaces induce sex-dimorphic secretion of angiogenic factors by gingiva-derived mesenchymal stromal cells

doi: 10.1016/j.jds.2025.09.013

Figure Lengend Snippet: Laser-modified titanium promotes the secretion of extracellular vesicles enriched with CCN1 and EDIL3 from human gingiva-derived mesenchymal stromal cells (GMSCs). (A) Primary GMSCs from one female and one male donor were cultured on machined, lasered, and SLA discs for 72 h, followed by ATAC-seq and RNA-seq analysis. Heatmaps show exosome-related genes. M, machined. L, lasered. S, SLA (sand-blasted, large-grit, acid-etched). (B) Primary GMSCs were cultured on machined, laser-treated, and SLA (sand-blasted, large-grit, acid-etched) titanium discs for 72 h. The conditioned medium was collected and centrifuged at 300× g for 10 min. The supernatant was then filtered through a 0.22 μm membrane, and particle size was analyzed using nanoparticle tracking analysis (NTA). Data are presented as mean values (n = 8 per group; 4 females and 4 males). (C) Western blot analysis was performed to evaluate CCN1, EDIL3 and ADM2 protein levels in cells, extracellular vehicles (EVs), and non-extracellular vesicle (non-EV) fractions derived from GMSCs (51-year-old male, 51M, and 63-year-old female, 63F) were cultured on machined (M), lasered (L), and SLA (sand-blasted, large-grit, acid-etched, S) titanium discs for 72 h. Laser-modified surfaces increased CCN1 in male GMSC EVs and EDIL3 in female EVs compared to machined surfaces. Lower panels show Ponceau S staining of the corresponding PVDF membranes. Results shown are representative of at least three independent experiments. CCN1, cellular communication network factor 1. ADM2, adrenomedullin 2. EDIL3, EGF like repeats and discoidin domains 3. TSG101, tumor susceptibility gene 101.

Article Snippet: Subsequently, the medium was replaced with fresh exosome-free α-MEM and cells were cultured for an additional 48 h. Conditioned medium was collected for the tube formation assay, and GMSCs were harvested for RNA extraction. mRNA expression was validated using TaqMan Fast Advanced Master Mix (4444556, Invitrogen) with TaqMan Gene Expression Assays (Invitrogen) for CYR61 (Hs00155479_m1), EDIL3 (Hs00964112_m1), and GAPDH (Hs02786624_g1), according to the manufacturer's protocol.

Techniques: Modification, Derivative Assay, Cell Culture, RNA Sequencing, Membrane, Western Blot, Staining

Journal: Journal of Dental Sciences

Article Title: Laser-modified titanium surfaces induce sex-dimorphic secretion of angiogenic factors by gingiva-derived mesenchymal stromal cells

doi: 10.1016/j.jds.2025.09.013

Figure Lengend Snippet: Silencing CCN1 or EDIL3 in gingiva-derived mesenchymal stromal cells (GMSCs) impairs HUVEC angiogenesis. GMSCs were seeded on machined, lasered, or SLA (sand-blasted, large-grit, acid-etched) titanium discs and transfected with siRNAs targeting CCN1 or EDIL3 for 24 h. (A) Relative mRNA expression of CCN1 and EDIL3 after siRNA transfection. NC, negative control siRNA. (B) Representative images of HUVEC tube formation in response to conditioned medium (CM) collected from female (F) and male (M) GMSCs. Images were captured using the ImageXpress Pico system. Scale bars = 500 μm. (C) Quantification of tube formation (branch points, total length, segments, and nodes) using ImageJ. Data are mean ± standard deviation (n = 2, one female and one male). One-way ANOVA analysis with Tukey's multiple comparison test. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. ns, not significant.

Article Snippet: Subsequently, the medium was replaced with fresh exosome-free α-MEM and cells were cultured for an additional 48 h. Conditioned medium was collected for the tube formation assay, and GMSCs were harvested for RNA extraction. mRNA expression was validated using TaqMan Fast Advanced Master Mix (4444556, Invitrogen) with TaqMan Gene Expression Assays (Invitrogen) for CYR61 (Hs00155479_m1), EDIL3 (Hs00964112_m1), and GAPDH (Hs02786624_g1), according to the manufacturer's protocol.

Techniques: Derivative Assay, Transfection, Expressing, Negative Control, Standard Deviation, Comparison

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A A diagram illustrating the setup of the experiment uxIRI. B Representative images of H&E staining, KIM1, F4/80, and CCN1 immunostaining in kidney tissues. C Renal CCN1 protein and mRNA expression. D Serum CCN1 levels in mice after I/R. n = 4 mice per group. E CCN1 expression in RTECs from AKI patients by single-cell RNA-seq. F Representative fluorescent micrographs and quantification analysis of CCN1 (yellow), AQP1 (red), and F4/80 (green) staining in I/R-AKI mice kidney. n = 4 mice per group. G CCN1 mRNA expression levels in HK-2 cells after H/R or H 2 O 2 stimulation. H CCN1 protein expression levels in HK-2 cells after H/R or H 2 O 2 stimulation. I Secreted CCN1 protein levels in culture supernatants of HK-2 cells after H/R or H 2 O 2 stimulation. n = 3 per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Staining, Immunostaining, Expressing, RNA Sequencing

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A A diagram illustrating the CCN1 protein administration in mice after I/R. B Blood urea nitrogen and serum creatinine levels in mice after I/R. C Representative images and quantification analysis of H&E staining, and KIM1 immunostaining in kidney tissues. D Density ridge plots showing the expression distributions of core-enriched gene sets identified by GSEA. E Selected enrichment plots from GSEA based on the gene enrichment profiles on CCN1-treated I/R kidneys versus I/R controls. F Gating strategy for CD45 + F4/80 + CD11b + cells in the kidney after I/R by flow cytometry. G Subsequent gating for macrophage polarization markers, including CD206, Ly6C, CD86 and CCR2 in the kidney after I/R by flow cytometry. n = 4 mice per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Staining, Immunostaining, Expressing, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A A diagram of uxIRI setup in a tubule-specific CCN1 knockdown model induced by renal pelvic AAV9-Ksp-shCCN1 injection. B Blood urea nitrogen and serum creatinine levels in mice after I/R. Representative images and quantification analysis of C H&E staining, D KIM1 and E F4/80 immunostaining in kidney tissues. F Gating strategy for CD45 + F4/80 + CD11b + cells in the kidney after I/R by flow cytometry. G Subsequent gating for macrophage polarization markers, including CD86, CCR2, Ly6C, and CD206 in the kidney after I/R by flow cytometry. n = 4 mice per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Knockdown, Injection, Staining, Immunostaining, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A A diagram of BMDMs isolation and CCN1 protein treatment. B , C Flow cytometry analysis of Ly6C⁺MHCII⁻ monocytes and Ly6C⁻MHCII⁺ macrophages following CCN1 stimulation. n = 3 per group. D CCK-8 assay of cell viability and adherence after CCN1 treatment. n = 6 per group. E Contour plot of CD86⁺, CCR2⁺, Ly6C⁻ and CD206⁺ BMDMs by flow cytometry. F Histogram of CD86, CCR2, Ly6C and CD206 of BMDMs by flow cytometry. n = 3 per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Isolation, Flow Cytometry, CCK-8 Assay

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A Volcano plot showing differentially expressed genes in BMDMs with or without CCN1 treatment under unstimulated or LPS and IFN-γ-stimulated conditions. B Heatmaps of marker genes of seven macrophage subpopulations for macrophage subpopulations. C Concentration of growth factors IGF-1 and PDGF-BB in BMDMs medium. D The mRNA expression of growth factors ( Igf1 , Pdgfb , Vegfa , Hgf , and Hbegf ) in BMDMs. E A diagram of the transwell co-culture system with CCN1-treated BMDMs in the top chamber and HK-2 cells in the bottom chamber. F Representative image of EdU immunofluorescence staining and quantification of EdU + proportion in HK-2 cells in the bottom chamber. G Flow cytometry analysis of EdU⁺ HK-2 cells in the co-culture system. n = 3 per group. H Representative images and quantitative analysis of Ki67 immunostaining in I/R mice kidney with AAV-shScr and AAV-shCCN1. I Representative images of Ki67 immunostaining in I/R mice kidney with CCN1 treatment and Clodronate Liposomes (CL). n = 4 mice per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Marker, Concentration Assay, Expressing, Co-Culture Assay, Immunofluorescence, Staining, Flow Cytometry, Immunostaining, Liposomes

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A BayesPrism workflow of scRNA-seq ( GSE174324 ) and bulk RNA-seq transcriptomic data integration and deconvolution based on BayesPrism to infer joint. B Cell-type fraction of deconvolution analysis across all samples. C Comparison of cell-type fractions following CCN1 treatment under normal and LPS-stimulated conditions. n = 4 per group. * P < 0.05. D , E The protein and mRNA expression of ARG1 in BMDMs. F , G Contour plot and histogram of flow cytometry of the proportion of ARG1 + cells in BMDMs. H The concentration of urea and Arginase activity in BMDMs. n = 4 per group. Data are presented as mean ± SD. I Representative fluorescent micrographs and quantification analysis of ARG1 (yellow), F4/80 (red) and EGFP-tag staining in I/R mice kidney with AAV-shScr and AAV-shCCN1 injection. J Flow cytometry of I/R mice kidney with CCN1 treatment. n = 4 mice per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: RNA Sequencing, Comparison, Expressing, Flow Cytometry, Concentration Assay, Activity Assay, Staining, Injection

Journal: Cell Death & Disease

Article Title: Tubule-derived CCN1 drives renal repair via α v β 5 -STAT6-ARG1-dependent reprogramming of macrophages

doi: 10.1038/s41419-025-08340-2

Figure Lengend Snippet: A GSEA enrichment of STAT-related pathways in CCN1-treated BMDMs. B Protein levels of p-STAT6, STAT6 and ARG1 in BMDMs. C Co-IP and MS workflow for CCN1-interacting proteins. D Overlap of CCN1-interacting proteins identified by Co-IP/MS and BioGRID database. E Molecular docking of CCN1 with ITGAV and ITGB5. F Co-IP validation of CCN1 binding to ITGAV and ITGB5 in BMDMs. G Protein levels of p-STAT6, STAT6 and ARG1 after α v β 5 integrin inhibitor treatment. n = 3 per group. Data are presented as mean ± SD.

Article Snippet: BMDMs were seeded in the top chamber and treated with 2 μg/mL CCN1 protein, 50 μM ARG1 inhibitor (Numidargistat, MedChemExpress), or a combination of both.

Techniques: Co-Immunoprecipitation Assay, Biomarker Discovery, Binding Assay

Journal: Molecular Therapy Oncology

Article Title: Targeted nanoencapsulation of tunicamycin reduces toxicity while improving its therapeutic effectiveness in pancreatic cancer cells

doi: 10.1016/j.omton.2025.201047

Figure Lengend Snippet: Effect of TM on 3D co-culture as in vitro desmoplasia and CCN1 expression in PDAC cells (A) In vitro desmoplasia assay using co-culture of untreated or NP TM-treated Panc-1 cells and fibroblast cells as depicted in upper panel. Arrow indicates a desmoplasia-like structure. Scale bars, 50 μm. (B) 3D co-culture of Panc-1 and pancreatic Stellate cells in True Gel3D hydrogel for the detection of tumor-stroma interaction and F-actin expression. Scale bars, 50 μm. (C) Immuno-western blot analysis of CCN1 in untreated, free, and nanoencapsulated TM-treated Panc-1 cell extracts. Cells were treated for 48 h, and 50 μg total protein/sample was separated in SDS-Page gel. (D) The graphs illustrated the signaling status of CCN1 in different samples derived from five independent experiments of each group. Data represent mean ± SD. The p value was calculated using the student t test .

Article Snippet: Anti-rabbit IgG (# 70745) was purchased from Cell Signaling, USA; anti-mouse IgG (# sc-516102) from Santa Cruz, Inc, USA; CCN1 polyclonal antibody (#26689-1-AP) and Neuropilin-1 (NRP-1) from ProteinTech, USA; and pSTAT3 polyclonal antibody (#9145) and STAT3 polyclonal antibody, peIF4E and EIF4E antibodies, and PCNA rabbit polyclonal antibody (#13110T) from Cell Signaling, Inc., USA.

Techniques: Co-Culture Assay, In Vitro, Expressing, Western Blot, SDS Page, Derivative Assay